Dubuis et al. - Source_Data_Files

Complete source data for Dubuis et al. contains two folders: 1. Folder "FC_Calibration" contains the source data used for calibration of the time-dependent furrow canal depth. It displays the chi_2 minimized furrow canal depth (delta_FC) for the 8 live imaged Drosophila embryos as function of time during n.c. 14 (see Materials and Methods). It also shows the mean furrow canal depth averaged over the 8 embryos (mean_delta_FC) and adjusted mean in fixed tissues (mean_delta_FC_fixed) that we used to estimate the age of the 163 fixed embryos. Column 1 contains the time in minutes after the onset of n.c. 14. Columns 2 to 9 contain the chi_2 minimized furrow canal depths (delta_FC_x, x being the embryo number) in for the 8 live imaged embryos. NaN means that we couldn't detect the furrow canal at that particular time. Column 10 contains the mean furrow canal depth averaged over the 8 embryos (mean_delta_FC) measured in micrometers. Column 11 contains the the furrow canal depth, measured in micrometers, which we used to estimate the age of fixed embryos. It was obtained by a 5% shrinkage of the previous column (see Materials and Methods). 2. Folder "Images" contains the original images of 201 Drosophila embryos at blastoderm stage immunostained against the four main gap genes (Kni, Kr, Gt, Hb). For each embryo with provide 5 12-bit images, each corresponding to a different optical channel (see Materials and methods). Channel 0 -- rat @ Kni (488nm); Channel 1 -- bright field (delta_FC); Channel 2 -- guinea pig @ Gt (568nm); Channel 3 -- rabbit @ Kr (594nm); Channel 4 -- mouse @ Hb (647nm). Images were taken with a Leica 20x HC PL APO NA 0.7 oil immersion objective, and with sequential excitation wavelengths of 488, 546, 594 and 633 nm. The bandwidth of the detection filters were set up as shown in Figure 1A to minimize fluorophore cross-talk while still allowing good detection in each optical channel. For each embryo, three high-resolution images (1024x1024 pixels, with 12 bits and at 100 Hz) were taken along the anterior-posterior axis (focused at the midsagittal plane) at 1.7 magnified zoom and averaged together. With these settings, the linear pixel dimension corresponds to 0.44 um.